Utrecht Sequencing Facility (USEQ)

About

The Utrecht Sequencing Facility provides support for Next Generation Sequencing (NGS) based research

We provide support for a broad range of services, including DNA/RNA isolation, sample preparation, Next Generation Sequencing, SNP fingerprinting, Spatial Omics and basic data analysis.

A comprehensive state-of-the-art infrastructure is available, including NGS sequencers, liquid handling robotics, bioinformatics tools and ICT.

You are welcome to take a look at our standard services or contact us for any customizations.

The Utrecht Sequencing Facility is sponsored by the University Medical Center Utrecht and the NWO infrastructure grant X-omics.

Contact us

Please contact us to discuss the possibilities for your research.

DNA / RNA isolation from cells, blood, tissue etc

Obtaining high quality sequencing data starts with high quality DNA and RNA. We use the Qiagen QiaSymphony SP for its proven robust high quality automated isolation of whole tissue, cells, organoids, blood. Standard services include human and mouse samples. Please contact us to discuss possibilities for samples of other species. For turn around times, see our FAQ on the USEQ website.

Sample requirements

We request samples to be submitted as either snapfrozen tissue, dry cell / organoid pellet, or dissolved in 200 ul ATL buffer (for DNA), dissolved in 400 ul RLT+ buffer (for RNA, stored at -80C).

Library Preparation

Convert your sample into sequence libraries

We facilitate the following library preparation methods using standard Illumina reagents. For turn around times, see our FAQ on the USEQ website.

Available kits:

  • TruSeq DNA Nano LT
  • TruSeq Stranded total RNA, ribo zero for human/mouse/rat
  • TruSeq Stranded mRNA polyA

A maximum of 24 (single index) or 96 (dual index) TruSeq indexes is available. Please contact us if you would like to use custom indexes. Bioinformatics demultiplexing service is supported for a restricted set of indexes.

Sample requirements

All concentrations should be measured with a fluorometric quantitation analysis (e.g. Qubit).

For DNA library preparation:

  • a minimum of 50 μl at a minimal concentration of 4 ng/μl.

For RNA library preparation:

  • RiboZero depletion: a minimum of 12 μl at a minimal concentration of 17 ng/μl.
  • PolyA enrichment: a minimum of 50 μl at a minimal concentration of 4 ng/μl.
  • RNA quality should be at least RIN 8 (RNA Integrity Number).
  • We strongly advise to include DNase treatment during the RNA isolation process.

Requester is responsible for collection of primary sample. Note that samples will be stored for a maximum of 2 months and subsequently discarded without further notice.

Next Generation Sequencing

USEQ operates a broad range of sequencing platforms, each with unique characteristics. Technical and cost characteristics determine the best fit for your experiment. Service fees include costs for chemicals, operator time, quality control, maintenance, lab information management system (LIMS) and troubleshooting.

Platform & Configurations

Currently, we operate the following Illumina platforms and configurations:

  • 1x NextSeq500
  • 2x NextSeq2000
  • 1x iSeq100
  • NovaSeq 6000 & X (external)
  • Nanopore MinIon
  • Nanopore GridIon X5
  • Nanopore PromethIon

Spatial Omics: Nanostring GeoMX & CosMX

Determine RNA or protein profiles based on morphology from small cell populations to within single cells

Digital Spatial Profiling: GeoMX

Current techniques offer a trade-off between spatial data and in-depth expression data. Bulk sequencing, for instance, generates in-depth data, but often it tells us little about where these cells were situated in the original tissue. Techniques like single molecule FISH and immunohistochemistry offer us spatial data, but only have a limited number of targets. USEQ offers a new technique called Nanostring GeoMX (Cell population based) & Nanostring CosMX (single cell or subcellular), which combines the spatial data with RNA or protein expression profiles. You provide the tissue of your choice and select regions that interest you. This can be done with Fresh Frozen or FFPE standard microscopy slides. Please call us for a detailed consult.